ORGANOIDS COMPRISING ISOLATED RENAL CELLS AND USE THEREOF
Described herein are organoids comprising admixtures of selected bioactive primary renal cells and a bioactive cell population, e.g., an endothelial cell populations, e.g. HUVEC cells, and methods of treating a subject in need thereof with such organoids. Further, the isolated renal cells, which may include tubular and erythropoietin {EPO}-producing kidney cell populations, and/or the endothelial cell populations may be of autologous, syngeneic, allogeneic or xenogeneic origin, or any combination thereof. Further provided are methods of treating a subject in need with the organoids.
1 . A method of forming an organoid comprising a heterogeneous renal cell population and a bioactive cell population, said method comprising culturing the heterogenerous renal cell population and a bioactive cell population in a culture system selected from the group consisting of i) 2D culture; ii) 3D culture: COL(I) gel; iii) 3D culture: Matrigel; iv) 3D culture: spinners, followed by COL(I)/Matrigel; and v) 3D culture: COL(IV) gel.
2 . The method of claim 1 wherein the heterogeneous renal cell population comprises a bioactive renal cell population.
3 . The method of claim 1 wherein the heterogeneous renal cell population comprises a B2 cell population comprising an enriched population of tubular cells, and wherein the heterogeneous renal cell population is depleted of a B1 cell population.
4 . The method of claim 3 wherein the heterogeneous renal cell population is further depleted of a B5 cell population.
5 . The method of claim 1 wherein the heterogeneous renal cell population comprises a cell population selected from B2, B2/B3, B2/B4, and B2/B3/B4.
6 . The method of claim 1 wherein the heterogeneous renal cell population comprises erythropoetin (EPO)-producing cells.
7 . The method of claim 1 wherein the bioactive cell population is an endothelial cell population.
8 . The method of claim 7 wherein the endothelial cell population is a cell line.
9 . The method of claim 7 wherein the endothelial cell population comprises HUVEC cells.
10 . The method of claim 1 wherein the cell populations are selected from xenogeneic, syngeneic, allogeneic, autologous and combinations thereof.
11 . The method of claim 1 wherein the heterogeneous renal cell population and the bioactive cell population are cultured separately for a first time period, combined and cultured for a second time period.
12 . The method of claim 11 wherein the second time period is at least 24 hours.
13 . The method of claim 12 wherein the second time period is 24 hours to 72 hours.
14 . The method of claim 1 wherein the renal cell population and bioactive cell population are at a ratio of 1:1.
15 . The method of claim 1 wherein the renal cell population and bioactive cell population are suspended in growth medium.
16 . An organoid made according the method of any one of claims 1 to 15 .
17 . An organoid comprising a heterogeneous renal cell population and a bioactive cell population.
18 . The organoid of claim 17 wherein the bioactive cell population is an endothelial cell population.
19 . The organoid of claim 18 wherein the heterogeneous renal cell population comprises a B2 cell population comprising an enriched population of tubular cells, and wherein the heterogeneous renal cell population is depleted of a B1 cell population.
20 . The organoid of claim 19 wherein the heterogeneous renal cell population is further depleted of a B5 cell population.
21 . The organoid of claim 17 wherein the heterogeneous renal cell population comprises a cell population selected from B2/B3, B2/B4, and B2/B3/B4.
22 . The organoid of claim 17 wherein the heterogeneous renal cell population comprises erythropoetin (EPO)-producing cells.
23 . The organoid of claim 18 wherein the endothelial cell population is a cell line.
24 . The organoid of claim 18 wherein the endothelial cell population comprises HUVEC cells.
25 . An injectable formulation comprising at least one organoid and a liquid medium.
26 . The formulation of claim 25 , wherein the liquid medium is selected from a cell growth medium, DPBS and combinations thereof.
27 . The formulation of claim 26 wherein the liquid medium is DPBS.
28 . The formulation of claim 25 , wherein the organoids are suspended in the liquid medium.
29 . An injectable formulation comprising organoids and a temperature-sensitive cell-stabilizing biomaterial that maintains
(i) a substantially solid state at about 8° C. or below, and
(ii) a substantially liquid state at about ambient temperature or above.
30 . The formulation of claim 29 , wherein the organoids comprise bioactive renal cells.
31 . The formulation of claim 30 , wherein the organoids further comprise HUVEC.
32 . The formulation of claim 29 , wherein the bioactive cells are substantially uniformly dispersed throughout the volume of the cell-stabilizing biomaterial.
33 . The formulation of claim 29 , wherein the biomaterial comprises a solid-to-liquid transitional state between about 8° C. and about ambient temperature or above.
34 . The formulation of claim 29 , wherein the substantially solid state is a gel state.
35 . The formulation of claim 29 , wherein the cell-stabilizing biomaterial comprises a hydrogel.
36 . The formulation of claim 35 , wherein the hydrogel comprises gelatin.
37 . The formulation of claim 36 , wherein the gelatin is present in the formulation at about 0.5% to about 1% (w/v).
38 . The formulation of claim 36 , wherein the gelatin is present in the formulation at about 0.75% (w/v).
39 . A method of treating kidney disease in a subject in need comprising administering at least one organoid comprising a heterogeneous renal cell population and a bioactive cell population.
40 . The method of claim 39 wherein the bioactive cell population is an endothelial cell population.
41 . The method of claim 39 wherein the heterogeneous renal cell population comprises a B2 cell population comprising an enriched population of tubular cells, and wherein the heterogeneous renal cell population is depleted of a B1 cell population.
42 . The method of claim 41 wherein the heterogeneous renal cell population is further depleted of a B5 cell population.
43 . The method of claim 39 wherein the heterogeneous renal cell population comprises a cell population selected from B2, B2/B3, B2/B4, and B2/B3/B4.
44 . The method of claim 39 wherein the heterogeneous renal cell population comprises erythropoetin (EPO)-producing cells.
45 . The method of claim 40 wherein the endothelial cell population is a cell line.
46 . The method of claim 40 wherein the endothelial cell population comprises HUVEC cells.
47 . A method of treating kidney disease in a subject in need comprising administering an injectable formulation according to any one of claims 25 - 38 .
48 . A method according to any one of claims 39 - 46 wherein the subject is a mammal.
49 . The method according to claim 48 wherein the mammal is a human.
50 . The method according to any one of claims 39 - 46 wherein the subject has a kidney disease.
51 . The method according to any one of claims 39 - 46 wherein an improvement in any one of the following measures of anemia (Hct, Hgb, RBC), inflammation (WBC), urine concentration (spGrav) and azotemia (BUN) is observed.
52 . Use of an organoid in the preparation of a medicament for the treatment of kidney disease.